摘要
目的:探讨三七总皂苷(PNS)对人脐静脉内皮细胞(HUVECs)过氧化氢(H2O2)所致损伤的保护作用。方法:采用体外培养HUVECs,建立过氧化氢(500μmol/L作用24h)内皮细胞损伤模型。分为4组:空白组、过氧化氢组、PNS低剂量(L)组(浓度为30μmol/L)、PNS高剂量(H)组(浓度为60μmol/L)。正常培养6h,用噻唑蓝(MTT)比色法测定HUVECs增殖及细胞活性,荧光素双醋酸酯(FDA)染色检测HUVECs生命力强度,蛋白免疫印迹(Western Blot)法检测各组细胞损伤相关蛋白超氧化物歧化酶(SOD)、丙二醛(MDA)、乳酸脱氢酶(LDH)、谷胱甘肽(GSH)的表达。用荧光定量(PCR)法检测各组细胞损伤相关基因SOD、MDA、LDH、GSH的表达。结果:PNS在一定浓度范围内可显著提高受损HUVECs活力,Western Blot结果表明SOD和GSH的蛋白表达升高,MDA和LDH的蛋白表达降低,差异具有统计学意义(P<0.01)。PCR结果与Western Blot结果相同。结论:PNS在一定浓度时可避免HUVECs不受H2O2的损伤。其分子作用机制可能是通过下调LDH、GSH蛋白表达,上调SOD、MDA蛋白表达而发挥作用。
Objective:To investigate the protective effect of panax notoginseng saponins(PNS)on human umbilical vein endothelial cells(HUVECs)induced by hydrogen peroxide(H2O2).Methods:HUVECs were cultured in vitro to establish the endothelial cell injury model of hydrogen peroxide(500 mol/L,24 h).The PNS at different concentrations were used for intervention.The cells were divided into four groups:Blank,Hydrogen Peroxide,and PNS Low(30 mol/L),PNS High(60 mol/L).Six hours later.With thiazole blue determined by MTT colorimetric method to determine HUVECs proliferation and cell activity.Using fluorescein diacetate(FDA)staining intensity of detection of HUVECs vitality.Western Blot method to detect each cell damage related protein superoxide dismutase(SOD),malondialdehyde(MDA),lactate dehydrogenase(LDH)and the expression of glutathione(GSH).By PCR method to detect each cell damage related genes SOD,MDA,LDH and GSH.Results:So,PNS could significantly increase the activity of the injured HUVECs within a certain concentration range.Western Blot results showed that the protein SOD and GSH were increased,while the protein MDA and LDH were decreased.PCR results showed the same as WB.Conclusion:PNS can prevent HUVECs from being damaged by H2O2.The mechanism may be related to down-regulation of LDH and GSH,up-regulation of SOD and MDA.
作者
谢鹏
刘金海
耿胜男
张萌萌
张会丽
Xie Peng;Liu Jinhai;Geng Shengnan;Zhang Mengmeng;Zhang Huili(School of Pharmacy and Chemical Engineering,Zhengzhou University of Industrial Technology,Zhengzhou,450000,China)
出处
《亚太传统医药》
2020年第9期33-37,共5页
Asia-Pacific Traditional Medicine
基金
郑州工业应用技术学院科研资助项目(2018YB023)
关键词
三七总皂苷
人脐静脉内皮细胞
动脉粥样硬化
Total Saponins of Panax Notoginseng
Human Umbilical Vein Endothelial Cells
Atherosclerosis
